DEVELOPMENT OF HELPER-DEPENDENT ADENOVIRAL VECTORS FOR THE EXPRESSION OF MUNC13 PROTEINS

Helper-Dependent Adenoviruses (HdAd) are the ideal viral vector choice for
expression of Munc13 proteins due to the large packaging capacity needed (~9.8 kb) for
the triple expression cassettes of Munc13, iCre, and EGFP. Development of a HdAd
vector for the expression of Munc13 gene family proteins allows for the rapid expression
of molecules located at the synaptic active zone critical for neurotransmitter release. The Munc13 HdAd viral vector can be used to express proteins of the munc13 gene family in the calyx of Held. This will allow for the elucidation of the molecular mechanisms
employed by the calyx of Held in the process of encoding the onset and modulation of
sound. Production of an HdAd vector for the expression of Munc13 proteins occurs in
five parts; cloning, transfection, amplification, purification and titering. After production
of the HdAd vector, Munc13 protein expression was analyzed by western blot.